Fig. 7

Immunochemical detection of pathological SOD1 species in the SOD1-ALS cases with anti-SOD1^int antibody. a Double immunofluorescence staining of the lumbar spinal cord section of the SOD1-ALS case with C111Y mutation (the case IV-6). The section was stained with rabbit anti-SOD1^int and mouse anti-SOD1 (clone 1G2, MBL) antibodies followed by Cy3-modified anti-rabbit (red) and FITC-modified anti-mouse (green) secondary antibodies, respectively. A merged image (yellow) was also shown. b Serial sections of the primary motor cortex of the SOD1-ALS case (the case IV-6) were immunostained with (left) anti-SOD1^int and (right) anti-SOD1 (clone 1G2, MBL) antibodies and visualized with DAB staining. c The lumbar spinal cord (ventral horn) of the case III-4, which exhibited exceptionally long disease duration (~69 years), was immunostained with anti-SOD1^int antibody. d, e SOD1 species recognized by d anti-SOD1^int and e anti-SOD1 (FL-154, Santa Cruz) antibodies were quantified in the soluble fraction of the homogenates of ventral and dorsal horn regions of thoractic spinal cord of human samples by sandwich ELISA. The sandwich ELISA was performed in the absence (−DTT) or presence (+DTT) of pre-treatment of the samples with 10 mM DTT. The SOD1-ALS cases III-4 and IV-6 and three non-ALS controls (Additional file 1: Table S1) were examined. The averages were shown as bars